GSM7712022: Adult LK LPS 16 hrs; Mus musculus; ATAC-seq
Cells were sorted into HBSS with 2% FBS and allowed to rest on ice for 1 hr prior to centrifugation. Cells were pelleted at 350 x g for 5 min at 4 degrees and resuspended in 0.04% BSA in PBS and pelleted again at 350 x g for 5 min at 4 degrees. Supernatant was removed, and the pelleted cells were lysed by adding 45 ul of chilled lysis buffer (10 mM Tris-HCl (pH 7.4), 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 0.1% NP-40 Substitute, 0.01% Digitonin, 1% BSA, 1 mM DTT, 1 U/ul RNase inhibitor) and pipetted gently 3 times before being incubated on ice for 3 minutes. Resulting nuclei were washed by adding 50 ul of chilled wash buffer (10 mM Tris-HCl (pH 7.4), 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 1% BSA, 1 mM DTT, 1 U/ul RNase inhibitor). Nuclei were pelleted at 500 x g for 10 min at 4 degrees and washed in diluted nuclei buffer (10X Genomics). Multiome GEM generation and library preparation was performed according to 10X Genomics protocol CG000338, targeting 5000 cell data recovery.